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Chitosan-heparin nanoparticles and acellular matrix-depleted biomaterials treated with the nanoparticles

A biomaterial and nanoparticle technology, applied in anticoagulant treatment, types of packaging items, special packaging items, etc., can solve the problems of endothelial cells and tissue replacement speed and degree not meeting the ideal requirements, and achieve long-term sustainable efficacy , stable particle performance, and the effect of improving biocompatibility

Active Publication Date: 2016-01-13
吴忠仕 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the early stage, the inventor of the present application used three decellularization agents, trinaton, trypsin and nuclease, to treat the bovine jugular vein (Bovine Jugular Vein Conduit, BJVC) with appropriate concentration, temperature and action time, and further cross-linked it by photo-oxidation technology. Enhanced tissue stability, good biomechanical properties, and superior anti-calcification properties, but at present, the product still faces the speed and degree of endothelial cell and tissue replacement that cannot meet the ideal requirements

Method used

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  • Chitosan-heparin nanoparticles and acellular matrix-depleted biomaterials treated with the nanoparticles
  • Chitosan-heparin nanoparticles and acellular matrix-depleted biomaterials treated with the nanoparticles
  • Chitosan-heparin nanoparticles and acellular matrix-depleted biomaterials treated with the nanoparticles

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Experimental program
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Effect test

Embodiment 1

[0040] Preparation of chitosan-heparin nanoparticles: improve the ion cross-linking method, the molecular weight of chitosan is controlled between 100kD-300kD, the concentration of chitosan solution is 1-4mg / ml, the heparin is low molecular weight heparin and unfractionated heparin, the concentration is 0.5 -2mg / ml, the temperature of the solution system is: 37°C, and the medium-speed magnetic stirring method is used for 20 minutes.

[0041]Results: By controlling the factors related to the preparation of nanoparticles, the particle size of chitosan-heparin nanoparticles was controlled between 50nm and 200nm, the range of PDI was between 0.110 and 0.185, and the Zata potential was between -20mv and +30mv. Nanoparticles can form a good core-shell structure with negative charges on the outermost layer according to needs, and the stable release time can reach more than 60 days after being further reinforced by EDC / NHS. (See figure 1 )

[0042] Conclusion: Chitosan-heparin nanop...

Embodiment 2

[0044] (1) Decellularization of bovine jugular vein-dye-mediated photooxidation treatment: the patent: used for pulmonary artery vascular repair or intermediate biological valve pipeline and its preparation method (patent number: ZL200510032124.9) for decellularization and dye-mediated The oxidation reaction has two steps.

[0045] (2) Nanoparticles are combined with bovine jugular veins through a two-step method. First, the nanoparticles and bovine jugular veins are physically adsorbed and self-assembled, and then chemically cross-linked by EDC / NHS: the bovine jugular veins are placed in 2 mg / ml nano In the particle solution, the self-assembly reaction time is 6 hours under the condition of a shaking speed of 80 rpm, and then chemical cross-linking and cross-linking are carried out for 4 hours to improve the bonding firmness of the nanoparticles and the material. The concentration of EDC is 200mmol / L, and the concentration of NHS is 60mmol / L.

[0046] RESULTS: The nanopartic...

Embodiment 3

[0049] VEGF binds to the nanoparticle-loaded bovine jugular vein obtained in Example 2. A 50-200ng / ml VEGFPBS ​​solution was prepared, and the bovine jugular vein loaded with nanoparticles was placed in the VEGF solution, and reacted below 25°C for 4 hours. Locally fixed VEGF was detected by immunohistochemical method, and the remaining VEGF content in the solution was detected by ELISA method.

[0050] Results: VEGF could specifically bind to heparin in bovine jugular vein loaded with nanoparticles.

[0051] Conclusion: VEGF can be locally immobilized and slowly released by nano drug loading.

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Abstract

The invention relates to a chitosan nano-material, particularly a nano-material formed with heparin. The invention further relates to a cell matrix-removed biomaterial. The invention aims at providing chitosan-heparin nano-particles suitable for specific binding of vascular endothelial growth factor (VEGF) and heparin and capable of binding with the cell matrix-removed biomaterial. The biomaterial is prepared by using an ion cross-linking method. The biological valve-possessed duct for pulmonary artery vessel restoration or reconstruction provided by the invention has good endothelialization performance, antithrombotic performance, anti-bacteria performance, and long-term persistent curative effect, and is beneficial for self-replacement of the biological valve-possessed duct.

Description

Technical field: [0001] The invention relates to a chitosan nano material, especially a nano material formed with heparin. [0002] The invention also relates to a decellularized matrix biomaterial. technical background: [0003] The application of nanotechnology in biological drug delivery has achieved good results, but it is very rare to apply nanomaterials locally to exert curative effects. Applying the nano-biological drug-loading system to the modification of decellularized matrix materials, taking advantage of its large drug-loading capacity, large specific surface area and controlled release characteristics, will help the long-term and stable performance of biologically active substances such as heparin and VEGF, and accelerate the development of bioactive substances in the body. The effect of tissue turnover. Heparin and VEGF can also be assembled on the surface of blood vessels by nano-layer-by-layer self-assembly technology. [0004] Chitosan is a natural polyme...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08L5/08C08L5/10C08J3/24A61L27/26A61L27/50A61L27/36A61L27/54A61L33/10A61L33/08A61L33/18
Inventor 谭琦吴忠仕
Owner 吴忠仕
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